白果内酯对大鼠栓塞后脑水肿模型脑内水通道蛋白9的影响Inhibition effect of bilobalide on the expression of aquaporin 9 in rat edema models
秦海明,宋福林,袁鹏程,翟兴雯,张新,游松
QIN Hai-ming1,2,SONG Fu-lin2,YUAN Peng-cheng3,ZHAI Xing-wen1,ZHANG Xin1
摘要(Abstract)：
目的探讨白果内酯(bilobalide,BB)对脑内水通道蛋白9(aquaporing,AQP9)的影响与其抗水肿作用的关系。方法利用Wistar大鼠持续性中动脉栓塞(permanent middle cerebral artery occlu-sion,pMCAO)制作左半球栓塞后水肿模型,通过脑水含量测定、梗塞面积测量、免疫组织化学染色、Real-time RT-PCR方法检测AQP9的mRNA和蛋白质的表达变化,对比栓塞对照组和BB预治疗组的差异,论证BB抗水肿的机制。结果正常组两侧脑含水量无差异。pMCAO24 h后,栓塞对照组和BB预治疗组脑水含量两侧均高于正常组,且栓塞侧明显高于栓塞对侧。BB预治疗组与栓塞对照组相比,两侧脑水含量均明显减低。pMCAO可引起大鼠皮质和白质中AQP9的大量表达。BB预治疗组较栓塞对照组胶质细胞和神经元细胞AQP9的表达出现明显减弱,特别是pMCAO 8 h和16 h时左侧白质及右侧皮质减低明显。pMCAO后AQP9的mRNA持续增加,16 h时达峰值,BB预治疗组明显降低了mRNA的合成。结论 BB通过抑制神经细胞中AQP9mRNA和蛋白质的合成机制脑水肿的形成。由于皮质中胶质细胞数量远大于神经元细胞,抑制AQP9合成的作用就更加明显。
Objective To study the influence of bilobalide(BB)to aquaporin 9(AQP9)and its anti-edema effect.Methods In this study,permanent middle cerebral artery occlusion(pMCAO)in adult Wistar rats was used as model.The anti-edema mechanism of BB was established through the measurements of the brain water content and the volume of the infarction,immunohistochemistry staining method,the research on the variation of AQP9 mRNA and protein using real-time RT-PCR method,and the study on the difference between bilobalide pretreatment group(10 mg · kg-1,i.p.)and the control group.Results The contralateral brain water contents of the normal control group were always stable.After pMCAO for 24 h,the contralateral brain water contents of both of the embolization control group and pretreatment control group with bilobalide were higher than that of the normal control group,and the side of embolization was obviously higher than the contralateral embolization.The contralateral brain water contents of pretreatment control group with bilobalide were decreased obviously compared with the embolization control group.pMCAO could induce the marked expression of AQP9 in the cortex and subcortical white matter of the rats.The expression of AQP9 of the glial cells and neurons of pretreatment control group with BB were lowered obviously compared with that of the embolization control group.In particular,the left white matter and the right cortex were decreased markedly during pMCAO for 8h and 16 h,respectively.After pMCAO,mRNA of AQP9 continuously increased,and reached peak value during 16 h.The synthesis of mRNA was decreased obviously for the pretreatment control group with BB.Conclusions These data suggest that BB attenuates brain edema formation due to reduced expression of mRNA of AQP9 in neuron and the proteinic synthesis.Because the number of the glial cells is far more than the neurons in the cortex,the action of inhibit AQP9 synthesis is much more significant.
关键词(KeyWords)：
白果内酯;持续性中动脉栓塞;水通道蛋白9;免疫组织化学染色;实时定量RT-PCR
bilobalide;pMCAO;AQP9;immunohistochemistry;real-time RT-PCR
基金项目(Foundation): 沈阳军区总医院青年基金资助项目(08Q-Y108)
作者(Author):
秦海明,宋福林,袁鹏程,翟兴雯,张新,游松
QIN Hai-ming1,2,SONG Fu-lin2,YUAN Peng-cheng3,ZHAI Xing-wen1,ZHANG Xin1
DOI: 10.14066/j.cnki.cn21-1349/r.2012.08.004
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